TY - JOUR
T1 - TAF11 Assembles the RISC Loading Complex to Enhance RNAi Efficiency
AU - Liang, Chunyang
AU - Wang, Yibing
AU - Murota, Yukiko
AU - Liu, Xiang
AU - Smith, Dean
AU - Siomi, Mikiko C.
AU - Liu, Qinghua
N1 - Funding Information:
We thank Drs. S.W. Ding and R. Carthew for generously sharing reagents; Drs. J. Albanesi, Y. Liu, H. Siomi, and J. Ma for helpful comments on the manuscript; and H. Liu, T. Liang, J. Lin, Y. Li, J. Chang, J. Hancock, L. Chen, and M. Buszczak for technical assistance. Q.L. is a W.A. “Tex” Moncrief Jr. Scholar in Medical Research. This work is supported by a Grant-in-Aid for Scientific Research (to M.C.S.); the World Premium Initiative from the Ministry of Education, Culture, Sports, Science, and Technology; and grants from the Welch foundation (I-1608), American Heart Association (13GRNT16270022), and NIH (GM111367) (to Q.L.).
Publisher Copyright:
© 2015 Elsevier Inc.
PY - 2015/9/3
Y1 - 2015/9/3
N2 - Assembly of the RNA-induced silencing complex (RISC) requires formation of the RISC loading complex (RLC), which contains the Dicer-2 (Dcr-2)-R2D2 complex and recruits duplex siRNA to Ago2 in Drosophila melanogaster. However, the precise composition and action mechanism of Drosophila RLC remain unclear. Here we identified the missing factor of RLC as TATA-binding protein-associated factor 11 (TAF11) by genetic screen. Although it is an annotated nuclear transcription factor, we found that TAF11 also associated with Dcr-2/R2D2 and localized to cytoplasmic D2 bodies. Consistent with defective RLC assembly in taf11-/- ovary extract, we reconstituted the RLC in vitro using the recombinant Dcr-2-R2D2 complex, TAF11, and duplex siRNA. Furthermore, we showed that TAF11 tetramer facilitates Dcr-2-R2D2 tetramerization to enhance siRNA binding and RISC loading activities. Together, our genetic and biochemical studies define the molecular nature of the Drosophila RLC and elucidate a cytoplasmic function of TAF11 in organizing RLC assembly to enhance RNAi efficiency. Liang et al. show that TAF11, an annotated nuclear transcription factor, is part of the Drosophila RISC loading complex (RLC). They provide evidence that TAF11 assembles the RLC by facilitating Dcr-2-R2D2 tetramerization to enhance RNAi efficiency.
AB - Assembly of the RNA-induced silencing complex (RISC) requires formation of the RISC loading complex (RLC), which contains the Dicer-2 (Dcr-2)-R2D2 complex and recruits duplex siRNA to Ago2 in Drosophila melanogaster. However, the precise composition and action mechanism of Drosophila RLC remain unclear. Here we identified the missing factor of RLC as TATA-binding protein-associated factor 11 (TAF11) by genetic screen. Although it is an annotated nuclear transcription factor, we found that TAF11 also associated with Dcr-2/R2D2 and localized to cytoplasmic D2 bodies. Consistent with defective RLC assembly in taf11-/- ovary extract, we reconstituted the RLC in vitro using the recombinant Dcr-2-R2D2 complex, TAF11, and duplex siRNA. Furthermore, we showed that TAF11 tetramer facilitates Dcr-2-R2D2 tetramerization to enhance siRNA binding and RISC loading activities. Together, our genetic and biochemical studies define the molecular nature of the Drosophila RLC and elucidate a cytoplasmic function of TAF11 in organizing RLC assembly to enhance RNAi efficiency. Liang et al. show that TAF11, an annotated nuclear transcription factor, is part of the Drosophila RISC loading complex (RLC). They provide evidence that TAF11 assembles the RLC by facilitating Dcr-2-R2D2 tetramerization to enhance RNAi efficiency.
UR - http://www.scopus.com/inward/record.url?scp=84940890444&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84940890444&partnerID=8YFLogxK
U2 - 10.1016/j.molcel.2015.07.006
DO - 10.1016/j.molcel.2015.07.006
M3 - Article
C2 - 26257286
AN - SCOPUS:84940890444
SN - 1097-2765
VL - 59
SP - 807
EP - 818
JO - Molecular cell
JF - Molecular cell
IS - 5
ER -